使用摩根-埃尔森法和新铜试剂法对透明质酸酶作用于透明质酸的动力学研究。
Kinetic investigation of the action of hyaluronidase on hyaluronan using the Morgan-Elson and neocuproine assays.
作者信息
Vercruysse K P, Lauwers A R, Demeester J M
机构信息
Laboratory of General Biochemistry, University of Ghent, Belgium.
出版信息
Biochem J. 1995 Aug 15;310 ( Pt 1)(Pt 1):55-9. doi: 10.1042/bj3100055.
Abstract
In this paper we describe kinetic investigations of the action of testicular hyaluronidase on hyaluronan. We have compared the use of two spectrophotometric assays, the first based on the Morgan-Elson reaction and the second on the neocuproine reaction. With the neocuproine reaction Km was found to be 0.46 mg/ml and Vmax to be 126 nmol l-1 s-1. Because of a low sensitivity and the production of interfering precipitates, the Morgan-Elson assay cannot be used for kinetic investigation of the enzyme. Furthermore this assay is prone to interference from compounds such as disodium cromoglycate, (+)-catechine, penicillamine, CaCl2 and acetate buffer.
摘要
在本文中，我们描述了睾丸透明质酸酶对透明质酸作用的动力学研究。我们比较了两种分光光度法的应用，第一种基于摩根-埃尔森反应，第二种基于新铜试剂反应。通过新铜试剂反应，发现米氏常数（Km）为0.46mg/ml，最大反应速率（Vmax）为126nmol l-1 s-1。由于灵敏度低且会产生干扰性沉淀，摩根-埃尔森测定法不能用于该酶的动力学研究。此外，该测定法容易受到诸如色甘酸二钠、（+）-儿茶素、青霉胺、氯化钙和醋酸盐缓冲液等化合物的干扰。
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